In-situ growth rate of Microcystis spp. and their growth-limiting factors: use of cellular RNA content

In 2003 and 2004, we conducted field investigations of a canal during the summer algal bloom to estimate the in-situ growth rate of Microcystis spp. and its limiting factors. Cellular RNA content (RNA/cell), determined by the real-time PCR method with a primer specific for amplification of Microcystis rRNA, was used as an index of in-situ growth rate. A Microcystis bloom was found in the canal in summer 2004 but not in summer 2003, because of its coldness. Corresponding to this, the average value of RNA/cell in 2004 was significantly higher than that in 2003. Water temperature, light intensity, and NO3 and PO4 concentrations were regarded as the factors limiting the in-situ growth rate of Microcystis in the canal, and their effects were quantified on the basis of laboratory experimental data. Effects of temperature and light intensity (photoinhibition by excessive photon flux density) were found to be important in limiting the growth rate, and more severe limitation was suggested in 2003. We then estimated the in-situ growth rate from the combined effect of these limiting factors. The estimated in-situ growth rates correlated significantly with RNA/cell in each year and in the combined (2003 + 2004) data. This agreement between our two different methods for estimation of in-situ growth rate suggests the validity of our approaches. This study was first field application of cellular RNA content as an index of algal growth rate in natural water samples.